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The preparation method of beauverum

A technology of beauveria and fusarium, applied in the field of industrial microorganisms, can solve the problems of unsuitability for industrial production, cumbersome operation steps, and few samples to be processed, and achieve the effect of long time-consuming change, strong controllability, and short process cycle

Active Publication Date: 2016-04-27
NCPC NEW DRUG RES & DEV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The three separation and purification methods mentioned above all have the same disadvantages: namely, the operation steps are cumbersome, the cycle is long, and the processing samples are few, which is not suitable for industrial production.

Method used

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  • The preparation method of beauverum
  • The preparation method of beauverum
  • The preparation method of beauverum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (a) Preparation of Fusarium LF061 seed solution

[0032] Prepare the seed medium according to the following proportions: YeastExtract 2g, Peptone 4g, glucose 4g, add distilled water to 200ml; adjust the pH value to 5.8, and sterilize at 121°C for 20min.

[0033] Fusarium LF061 was inoculated on the seed medium, and cultured at 28° C. at 120 rpm for 48 hours to obtain seed liquid.

[0034] (b) Fusarium LF061 seed solution was fermented to produce beauveria:

[0035]Preparation of liquid fermentation medium: glucose 5g, soluble starch 20g, industrial peptone 30g, potassium chloride 0.1g, potassium dihydrogen phosphate 3g, sodium nitrate 1g, magnesium sulfate heptahydrate 0.5g, calcium carbonate 1g, add water to 1000ml, Natural pH value, sterilized at 121°C for 30 minutes;

[0036] The Fusarium LF061 seed solution was inoculated into the liquid fermentation medium with an inoculation amount of 15% by volume, and fermented in a shake flask at a fermentation temperature of...

Embodiment 2

[0039] (a) Preparation of Fusarium LF061 seed solution

[0040] Prepare the seed medium according to the following ratio: YeastExtract 20g, Peptone 40g, glucose 40g, add distilled water to make the volume to 2000ml; adjust the pH value to 5.3, and sterilize at 121°C for 20min.

[0041] Fusarium LF061 was inoculated on the seed medium, and cultured at 30° C. at 130 rpm for 72 hours to obtain seed liquid.

[0042] (b) Preparation of beauveria from Fusarium LF061 seed solution by fermentation

[0043] Preparation of liquid fermentation medium: glucose 300g, soluble starch 900g, industrial peptone 1500g, potassium chloride 6g, potassium dihydrogen phosphate 150g, sodium nitrate 45g, magnesium sulfate heptahydrate 3g, calcium carbonate 60g, add water to 30L, pH Naturally, sterilize at 121°C for 30 minutes;

[0044] The Fusarium LF061 seed solution was inoculated into the liquid fermentation medium with an inoculation amount of 5% by volume, and fermented in a 50L fermenter at a f...

Embodiment 3

[0046] Take 10 L of the beauverian fermentation broth prepared in Example 2, and the fermentation unit is 425 μg / mL. The beauverum fermented liquid was filtered to obtain 2.5 kg of mycelia. Add 5.0 L of methanol to the above-mentioned mycelia, stir and extract for 4 hours, then filter, and collect the beauveriacin extract. The extract was concentrated at 40°C with a vacuum of 0.09 MPa until no liquid flowed out, and the crude extract of beauveria was obtained with a purity of 53% (see attached figure 1 ). Take 10 g of the crude extract of beauveria, dissolve it in 60 mL of methanol, add the sample to a medium-pressure chromatography column (Ф5.5×50 cm) of UniPS30-300 polymer microspheres with a capacity of 1000 mL, and use 80% by volume of Methanol aqueous solution was used as mobile phase to wash 2000mL, then washed with methanol, and received in separate tubes with a flow rate of 1500mL / h. According to HPLC detection, the eluents with a purity greater than 95.0% of beauv...

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Abstract

The invention discloses a preparation method of beauvericin. The method comprises the steps of 1a, preparation of a fusarium LF061 seed solution preparation:inoculating a slant culture or spore liquid of strain LF061 into a seed culture medium, and culturing for 48-72 hours with a rotating speed of 120-160rppm at 28-32 DEG C to obtain the seed solution; 1b, preparation of beauvericin from the fusarium LF061 seed solution through fermentation: inoculating the seed solution of which the inoculation amount is 5-15% in percentage by volume into a liquid fermentation culture medium, wherein the fermentation temperature is 28-32 DEG C, the rotating speed is 120-160rpm, the ventilation volume is 10-30L / min, and the fermentation time is 96-144 hours; separating and refining obtained fermentation liquid to prepare beauvericin. According to the method disclosed by the invention, the beauvericin is produced through liquid fermentation, and the prepared beauvericin is high in purity and recovery rate and adaptable in industrial production.

Description

technical field [0001] The invention belongs to the technical field of industrial microorganisms, and in particular relates to a preparation method of beauveria. Background technique [0002] Beauvericin (BEA) is derived from the fungus Beauvericin ( Beauverlabassiana ) a six-membered cyclic desipeptide compound extracted and isolated from fermented mycelia, consisting of 3 N-methylphenylalanine and 3 (D)- α -Hydroxyisovaleric acid residues are alternately connected, and the molecular formula is C 45 h 57 N 3 o 9 , the structure is shown in formula (I). Beauveriacin is in the form of white powder, not easily soluble in water, but soluble in organic solvents such as methanol and acetone. The detectable ultraviolet absorption spectrum ranges from 204nm to 225nm. [0003] [0004] Beauverum is a lipopeptide compound with biological activities such as antibacterial, insecticidal, antiviral, anticancer, inhibition of cholesterol acyltransferase activity, and induction of...

Claims

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Application Information

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IPC IPC(8): C12P21/04C07K1/16C12R1/77
Inventor 段宝玲张立新张雪霞刘梅任风芝贺伟李宁白超弦李晓露贾啸静陈书红米贯东李丽红王秀琴董爱华王海燕段向东
Owner NCPC NEW DRUG RES & DEV
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